CN105646752B - A kind of Fluorescent star-like polymer and preparation method and application - Google Patents
A kind of Fluorescent star-like polymer and preparation method and application Download PDFInfo
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Abstract
Prepared the present invention relates to compound and protein molecular carrier, specifically disclose a kind of Fluorescent star-like polymer and preparation method and application.Fluorescent star-like polymer of the present invention has prepared the Fluorescent star-like polymer of different algebraically, carrying amino acid characteristics group using " divergent method " using perylene analog derivative and the like as fluorescent core.Such Fluorescent star-like polymer molecule, because its periphery carries special amino acid group, easily by live body culture cell recognition and swallows.And by non-covalent bond, it can be combined respectively with the protein molecular with negative electrical charge or positive charge, the stable complex of each self-forming.Fluorescent star-like polymer of the present invention can be used as carrier; it can protect and deliver foreign protein molecule and be successfully entered live body culture cell; pass through the method for fed diets; desinsection toxic protein molecule can efficiently enter cell in living insects body by the delivery function of polymer; facilitate it to play toxicity action, cause insect growth slow and be poisoned lethal.
Description
Technical field
The present invention relates to compound prepare and protein carrier field, specifically, be related to a kind of Fluorescent star-like polymer and
Its preparation method and application.
Background technology
Due to current large-scale plantation Bt genetically modified crops, Bt class chemical pesticides are abused without planning of science activities so that former
First responsive type insect sharply increases to Bt resistance, a great problem as field of pest control.By the method for science come
Breaking insect-resistant becomes particularly important.Conventional strategy is mainly the layout by science at present, in Bt genetically modified crops week
Plantation non-transgenic crop is enclosed to provide " sanctuary " of insect, or by increasing the diversity of Bt toxalbumin molecules, it is to avoid elder brother
The resistance gene frequency of worm expands rapidly, so as to delay the Bt resistances of insect.These methods can delay elder brother to a certain extent
The resistant evolution of worm, but be not easy in application process to implement.The practical problems such as quality, the planting scale of grower, limitation
Their application.After to a part of reason of resistance of toxalbumin molecule being insect genes mutation there are some researches show, insect,
Toxalbumin molecular receptor on intestinal cell film becomes XOR expression quantity and is greatly decreased so that external source toxalbumin molecule is difficult with reference to upper
Cell-membrane receptor, causes toxalbumin virulence to decline, and insect develops immunity to drugs.
Development of the high polymer material in protein molecular carrier system in recent years is very rapid, wherein protein molecular carrier body
Owner will include degradable polymer microspheres, hydrogel, multi-walled carbon nanotube, charge reversal type micella etc..And with fluorescence star
Polymer was not reported also as the research of protein molecular carrier.Perylene diimide class compound is a class light, heat, chemical stability
Very good fluorescent chemicals, fluorescence quantum efficiency is very high (close to 1).By its superior dyeability, always by
It is widely used in the photoelectric fields such as organic electro-optic device, laser dye.
In recent years, such compound starts to be applied to biological field, such as protein molecular targeting mark and cell-specific mark
Note etc..But poorly water-soluble this shortcoming, it directly affects its application in this field.Therefore the water-soluble of this kind of compound is improved
Property and enhancing and protein molecular compatibility, will to research and development novel protein molecular vehicle have very important significance.Amino acid
It is the basic component units of protein molecular, and polymerize obtained polyaminoacid and possess similar functional group to protein molecular, root
According to " similar compatibility " principle, both have extraordinary compatibility.Therefore water miscible amino acid group Dui perylenes class chemical combination is passed through
Beyond the region of objective existence, which encloses, to be modified, and can not only have been improved its water solubility but also can have been strengthened the compatibility to protein molecular.
Star polymer is simplest a kind of topological structure in dendrimers, convenient with design, and synthesis cycle is short
The characteristics of.In addition, star polymer also has some other advantage, such as intramolecule has cavity structure, functional group densities high
Deng.These advantages make it close into perylene class polyaminoacid and applied to possessing huge advantage in terms of protein molecular carrier setting meter.Cause
And, inventing a kind of simple, efficient method is used to prepare amino acid functionalised perylenes class Fluorescent star-like polymer, studies this kind of glimmering
Light star polymer, in the application in biomedicine field and insect control field, will have extremely important as protein molecular carrier
Meaning.
Publication No. CN103172776A Chinese patent discloses a kind of Fluorescent star-like polymer and carried as nucleic acid substances
The method that body is applied to live body culture cell and insect.Using perylene analog derivative and the like as fluorescent core, using " diverging
Method " has prepared different algebraically, has carried the Fluorescent star-like polymer of cation group.Such Fluorescent star-like polymer has fine
Biocompatibility and very low cytotoxicity, molecule periphery carries positively charged amino, can enter live body culture cell,
And it can be combined with negatively charged DNA or RNA, form a stable complex, carried as carrier and protect external source
Nucleic acid enters live body culture cell.The mixture of such carrier and DNA or RNA is injected by fed diets or body cavity, such load
Body can quickly carry DNA or RNA into cell in living insects body, being easy to these nucleic acid to play a role, interference insect target
The normal expression of gene is marked, so as to cause insect growth dysplasia until sick or death.
However, the Fluorescent star-like polymer in above-mentioned technical proposal belongs to amino polymer, the poly- of cationic is used as
Compound, amino polymer can utilize the electrostatic adsorption between DNA, reach the purpose of passenger gene, at present they
Can only as elecrtonegativity nucleic acid substances such as DNA and dsRNA carrier, and universal protein molecular carrier can not be used as.
The content of the invention
In order to solve problems of the prior art, protein molecular can be carried it is an object of the invention to provide a kind of
Fluorescent star-like polymer and preparation method and application.
In order to realize the object of the invention, present invention firstly provides a kind of Fluorescent star-like polymer, its structure such as formula (IV) institute
Show：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number, n=1~100, k=1~4, m=1~3.
Wave therein represents the chemical constitution that certain plays connection function.
Preferably, in described Fluorescent star-like polymer, n=5~30, k=1~2, m=2~3, R is carbon number 2~
Alkyl chain between 4 has more preferable effect.The reason is that the fluorescent core in the middle of as m=2-3 is by two or three naphthalene nucleus phases
Company forms conjugated structure , i.e. perylene and terylene.Their fluorescence emission wavelengths>600nm, higher than the ripple of biological context fluorescence
Long scope (400-550nm), the advantage for being conducive to it to play bio-imaging.As n=5~30, k=1~2, the weight of polymer
Multiple unit number is less, and sum is less than 1200, and the volume of polymer is relatively small, and specific surface area is relatively large, is conducive to egg
The absorption of white matter, and it is also relatively small with the compound volume of protein formation, it is advantageously implemented the phagocytosis of cell.And work as R
During for alkyl between carbon number 2~4, the water solubility of polymer was both ensure that, the intermolecular work between protein is added again
Firmly.
In one particular embodiment of the present invention, described Fluorescent star-like polymer is poly- ILE metering system
Acid amides, shown in chemical structural formula such as formula (V)：
Wherein, n=15.
Many benzene ring structure , that above-mentioned polymer has conjugation Ji perylene, and its fluorescence emission wavelengths is in 620nm or so.Polymer
Containing four polymer chains, molecular structure is the star structure of diverging, and each contains the residual of isoleucine in repeat unit
Base, i.e. carboxyl and alkyl structure.
Present invention also offers the preparation method of the Fluorescent star-like polymer, it with compound 2 is monomer that it, which is, with chemical combination
Thing 3 is initiator, and Fluorescent star-like polymer is obtained using ATRP living polymerizations；
Shown in the structure of monomeric compound 2 such as formula (II)：
Shown in the structure of compound 3 such as formula (III)：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number, k=1~4, m=1~3.
Compound in formula (III) has many benzene ring structures of conjugation, can be sent in the case where wavelength is 200-700nm illumination
Wavelength is 400-750nm fluorescence, and it is with multiple bromine isobutyl structures outside.This kind of compound can as ATRP initiators,
R is hydrogen atom or the alkyl for being 1~9 containing carbon number in monomeric compound in initiation formula (II), formula.Obtained polymer is kept
Contain a large amount of amino acid residues in fluorescent characteristic, and its peripheral polymer chain, whole polymer in diverging star structure.
More specifically, the preparation method is：The consumption of the versus compound 3 of compound 2 is 10~500 times, 60~
Reacted under conditions of 70 DEG C.Under this condition, the reaction can be obtained more in terms of reaction rate and molecular weight distribution
Good control.
Preferably, compound 3 is dissolved in organic solvent so that the concentration of compound 3 in the range of 2~5g/L, plus
Enter after the CuBr with the amount of the grade material of compound 3, carry out 3~4 freezings-pump drainage circulation；Afterwards under nitrogen protection, 5 are added
The part PMDETA (N, N, N', N, ' N "-five methyl diethylentriamine) of the amount of~15 times of materials of compound 3 and 10~500
The monomeric compound 2 of the amount of times material of compound 3；Again after freezing pump drainage, stir 10~15 minutes, make CuBr complete with part
Full complexing, reacts 30-120 minutes at 60~70 DEG C；After reaction terminates, solvent is removed, is dialysed, freeze-drying.
Preferably, the organic solvent is the mixture of butanone and methanol, the volume ratio of butanone and methanol is 2:1.This
Apply for that the ATRP polymerization used belongs to polymerisation in solution, using the mixture of butanone and methanol can realize oil-soluble initiator and
Water miscible monomer is dissolved in same system, so as to be conducive to the progress of polymerisation.
Further, the preparation method of the monomeric compound 2 is：Compound 1 is dissolved in sodium hydrate aqueous solution,
So that the pH value of solution is more than 10, then near 0 DEG C, methacrylic chloride is added dropwise, and supplement corresponding hydroxide
Sodium solution is to maintain pH value between 9~11；After completion of dropping, 1~3 hour is stirred at room temperature, afterwards with watery hydrochloric acid pair
Solution is acidified so that pH value is down between 1~3；It is extracted with ethyl acetate, most of solvent is removed with Rotary Evaporators,
Recrystallized by hot saturated solution method, obtain compound 2；
Shown in the structure of compound 1 such as formula (I)：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number.
The present invention still further provides the Fluorescent star-like polymer as the application of protein molecular carrier.
It is described application specifically, by Fluorescent star-like polymer and foreign protein molecular mixing formation complex, then with carefully
Born of the same parents are incubated altogether, the Fluorescent star-like polymer carrier for carrying foreign protein molecule is entered intracellular by endocytosis,
Or the method by injecting or being fed with insect, the Fluorescent star-like polymer carrier for carrying foreign protein molecule is entered insect bodies
It is interior.
Optionally, the Fluorescent star-like polymer is applied to active somatic cell：
Such Fluorescent star-like polymer is added in cell culture fluid with certain concentration, during one section of active somatic cell of culture
Between after, such compound runs through endocytosis and enters intracellular, by fluorescence microscope photographic analysis, clear observation
The distribution situation of such compound molecule in the cell.Such compound foreign protein molecule is in vitro mixed to form compound
Body, is then incubated after a period of time with the above method together with cell, and such compound plays protein molecular function vector, carries
Foreign protein molecule rapidly enters cell, so as to external source functional protein molecule function in the cell.
Optionally, the Fluorescent star-like polymer is applied to living insects：
Such Fluorescent star-like polymer and functional exogenous desinsection Bt toxalbumin molecular mixing formation complex, with certain
Concentration be added in insect feedstuff, for insect's food-taking for a period of time after, such compound play protein molecular function vector,
Protection external source toxalbumin molecule readily penetrates through funnel barrier in insect gut, into enterocyte, is added significantly to enter target
The protein molecular quantity of cell is marked, with few consumption, toxic action of the protein molecular to insect is realized so that insect growth
It is slow and be poisoned to death.The process stands good in the resistant insects for lacking the toxalbumin molecular receptor, breaks
Insect expands the insecticidal spectrum of Bt toxalbumin molecules to the resistance of Bt toxalbumin molecules.
As a kind of new protein molecular carrier that can be followed the trail of by fluorescence, such polymer is delivered in pharmaceutical protein molecule
Biomedical sector and field of pest control there is good application value.
The present invention is on the basis of there is provided Fluorescent star-like polymer, it is further provided a kind of malicious insecticidal method, its
It is, using foregoing Fluorescent star-like polymer as protein molecular carrier, to carry external source toxalbumin molecule and enter intracellular or insect bodies
It is interior, so as to kill insect.
Optionally, the Fluorescent star-like polymer carrier can inject or feeding insect by way of, into insect bodies, from
And kill insect.
Preferably, the foreign protein molecule is represented as the histone of positive charge or the Bt toxalbumin of negative electrical charge, should
The Fluorescent star-like polymer that albuminoid is provided with the present invention can be combined preferably, enter cell by efficiently delivery.
The beneficial effects of the present invention are：
The invention provides the new and effective protein carrier of a class, toxic protein molecule can be transported, breaks external source toxalbumin
The barrier of the entrance insect enterocyte of molecule, greatly improves the insecticide efficiency of toxalbumin particularly Bt toxalbumin, expands the malicious eggs of Bt
White insecticidal spectrum.
The present invention has synthesized using organic fluorescence chromophore as core and carried the work(of amino acid group by MOLECULE DESIGN
Can property Fluorescent star-like polymer.Such polymer has good light, heat, chemical stability, structure designability, excellent life
The characteristics such as thing compatibility.
Step of the present invention is simple and easy to apply, can the programmable star polymer of preparation structure rapidly：On the one hand can be by changing
The algebraically of initiator, controls the arm number of polymer；On the other hand polymer can be regulated and controled by adjusting feed ratio or reaction time
Chain length.
There are a large amount of amino acid residues, carboxyl therein in the architectural feature for the Fluorescent star-like polymer that the present invention is prepared
Condition is provided for the absorption of protein so that either electronegative or electropositive protein can pass through with alkyl
Electrostatic Absorption or hydrogen bond and Van der Waals force and polymer formation stable bond.Formed importantly, having adsorbed after protein
Compound can smoothly enter cell, this has very big contact with amino acid residue.Specifically, what the present invention was provided is glimmering
Photostar shape polymer molecule periphery carries amino acid group, can enter living cells, and can pass through ionic bond and positively charged
The protein molecular of lotus such as histone molecule is combined, and forms a stable compound, can be protected simultaneously as protein molecular carrier
Carry foreign protein molecule and enter living cells.Such carrier molecule also can be by Van der Waals force or hydrogen bond action, with reference to negatively charged
Bt toxalbumin molecules, delivery Bt protein moleculars enter insect enterocyte, trigger lethal processes, make insect growth hypoevolutism and
It is poisoned to death.Therefore, such carrier as a kind of new fluorescence polymer protein molecular carrier in biomedicine field and agriculture
Industry insect control field has good application value.
Brief description of the drawings
Fig. 1 is the route map of synthesis ILE Methacrylamide
Fig. 2 is 4Br-PDI synthetic route chart
Fig. 3 is the synthetic route chart of poly- ILE Methacrylamide
Fig. 4 is the nuclear magnetic spectrogram of poly- ILE Methacrylamide
Fig. 5 is the UV absorption and fluorescence spectra of the poly- ILE methacryl amine aqueous solution of various concentrations
Fig. 6 is the UV absorption and fluorescence that poly- ILE Methacrylamide and histone molecule (HS) are had an effect
Spectrogram
Fig. 7 is that carrier molecule carrying foreign protein molecule enters cell.A ' carriers carry foreign protein molecule and enter cell
Fluorescence imaging figure.B ' are carried into the protein molecular fluorogram of cell.B " enter the carrier fluorescence figure of cell.D. it is different
The entrained foreign protein molecular fluorescence intensity for entering cell of incubation time carrier molecule.
Fig. 8 is that carrier/Bt toxalbumin molecule prevents and treats anti-Bt insects.A. the insect larvae of independent carrier is fed.B. feed single
The insect larvae of only toxalbumin molecule grows normally.C. larval growth is sent out after feeding carrier/Bt toxalbumin molecular complexes
Educate and be obstructed.D. dead larvae after carrier/Bt toxalbumin molecular complexes is fed.
Embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this area
Art personnel can carry out various modifications and replacement in the case of without departing substantially from spirit of the invention and spirit to the present invention.
Embodiment 1
Embodiment 1 is by taking the poly- ILE Methacrylamide of star as an example, for illustrating fluorescence star of the present invention
The preparation method of polymer.
(1) synthesis of monomer：As shown in Figure 1, ILE (1.0g, 7.5mmol) is dissolved in sodium hydroxide water
(1mol/L, 7.5mL) and stirred in solution.Methacrylic chloride (1.17g, 11.2mmol) is added dropwise dropwise at 0 DEG C,
The sodium hydrate aqueous solution for being 2mol/L with concentration during the dropwise addition of 30 minutes adjusts pH value, it is maintained near 10.Return
When starting reaction to room temperature (25 DEG C), pH value is maintained near 7.5.After 1 hour, carried out with aqueous hydrochloric acid solution (2mol/L)
Acidifying, is then extracted (4 × 30mL) with ethyl acetate.Organic phase uses Rotary Evaporators after anhydrous sodium sulfate drying
Most of solvent is removed, product is obtained by the method for recrystallization afterwards and finally gives product 1.4g, i.e. ILE methyl
Acrylamide, yield 90%.
(2) 4Br-PDI synthesis：As shown in Figure 2, by Si Qiang Ji perylene diimides (4OH-PDI, 400mg, 0.32mmol)
40mL THF are dissolved in, 250mL three-necked flasks are transferred to, lead to nitrogen 15 minutes, air is excluded, flask is added in ice bath, until
Cool and be maintained near 0 DEG C, first add triethylamine (1.80mL, 12.8mmol), 2- bromine isobutyl acylbromides are then added dropwise dropwise
(1.60mL, 12.8mmol).After stirring 10 minutes, it is gradually heating to room temperature and starts reaction.Detected and reacted using thin-layer chromatography
Degree, 48 as a child terminate reaction.The method being washed with water removes the salt in reactant, obtains crude product.Use dichloromethane
Eluent is done, pillar layer separation obtains pure products, i.e. Si Xiu perylene diimides (4Br-PDI) 450mg, yield 76%.
(3) synthesis of poly- ILE Methacrylamide：As shown in Figure 3, by initiator 4Br-PDI (10.0mg,
5.4 μm of ol) and catalyst CuBr (3.1mg, 21.6 μm of ol) add 25mL reaction tubes in, with the mixing of 2mL 2- butanone and methanol
Thing (volume ratio 2:1) dissolve, the purpose of deoxygenation is reached by three freezing pump drainage processes, adds and matches somebody with somebody under the protection of inert gas
Body PMDETA (N, N, N', N, ' μm ol of N "-five methyl diethylentriamine, 77.8mg, 450) and ILE metering system
Acid amides (430mg, 2.16mmol), then with rubber stopper seal, after stirring CuBr and part complexing completely, by reaction tube plus
Enter and start reaction in 65 DEG C of oil baths.The polymer of respective chain length is can obtain by the specific time：As 15 chains can be obtained within 30 minutes
Long polymer.Reaction terminate after, by rotary evaporation remove solvent, by residue dialyse three days, change within every 5 hours once go from
Sub- water, eventually passes freeze-drying and obtains the poly- ILE Methacrylamide of final product, abbreviation P1, its nuclear magnetic spectrogram is shown in
Accompanying drawing 4.
The optical characteristics of the poly- ILE Methacrylamide of experimental example 1
The poly- ILE Methacrylamides (P1) of 1.2mg are dissolved in 8mL pure water, be made concentration for 1.25 ×
10-5The mol/L aqueous solution, it is 6.25 × 10 that concentration is diluted to respectively-6Mol/L, 4.16 × 10-6Mol/L, 2.76 × 10- 6Mol/L and 1.84 × 10-6The mol/L aqueous solution, and their UV absorption and fluorescent emission spectrogram are tested respectively, see accompanying drawing
5。
Influence for observing protein to P1 photoluminescent properties, it is 5.00 × 10 to take concentration-5Mol/L Histone (HS)
It is 1.84 × 10 that the μ L of the aqueous solution 20, which are added drop-wise to concentration,-6In the mol/L P1 aqueous solution, i.e., portion HS is added in the P1 aqueous solution, surveyed
Try and record its fluorescence spectrum, and add 8 parts of HS of total successively in the same manner, obtained fluorescence spectrum, which compares, sees accompanying drawing
6。
The delivery of the foreign protein molecule of experimental example 2 enters living cells
The amino star polymer carrier molecule of synthesis is combined with external source basic protein molecules formation carrier/protein molecular
Thing, is incubated together with live body culture cell, has investigated the ratio of protein molecular and carrier complexing, i.e. carrier molecule institute band anion
The quantity of electric charge and protein molecular influence of the quantity of electric charge ratio (N/P) to cell transfecting with cation.As shown in Figure 7, pass through
Research is with different N/P ratios (4：1,2：1,1:1；2：Isosorbide-5-Nitrae：1) carrier molecule and protein molecular compound transfectional cell formed
As a result find, carrier molecule shows good carrying capacity under the conditions of all N/P ratios.With 4：1 N/P synthetic vectors/egg
It is intracellular during 0.5h to can detect that foreign protein molecular signal during white molecular complex incubated cell, show that carrier molecule can
Quickly and efficiently carry foreign protein molecule and enter living cells.
Experimental example 3 prevents and treats insect with the synergy of insecticidal proteins molecule
Carrier molecule combination Bt toxalbumin molecule kills the insect resistant to Bt
Under the conditions of 25 DEG C, by 4 μ g carrier molecules and 1 μ g disinsection Bt proteins molecular mixing formation carrier/Bt toxalbumin point
Sub- compound, after half an hour, adds in 2mg insect feedstuffs, the first-instar young just hatched is individually fed after being mixed with feed.Often
Taken pictures every 24h observations, and measure its body length and body weight.2 control groups are set herein, are respectively：Independent Bt toxalbumin molecules, individually
Carrier molecule.After 72h, the larva of carrier molecule/Bt toxalbumin molecule feeding is poisoned to death.After 12d, the children survived is observed
Worm, it has been found that the larvae development of carrier molecule/Bt toxalbumin molecular complex feeding is slow, and body length compares remaining with body weight
Control has decline.Result above is shown in accompanying drawing 8.Therefore, carrier molecule combination Bt toxalbumin molecule enters in the enterocyte of larva,
The effect of Bt toxalbumin molecules is given full play to, suppresses insect and normally grows, makes insect poisoning lethal.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (12)
1. a kind of Fluorescent star-like polymer, it is characterised in that shown in its structure such as formula (IV)：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number, n=1~100, k=1~4, m=1~3.
2. Fluorescent star-like polymer according to claim 1, it is characterised in that n=5~30, k=1~2, m=2~3, R
The alkyl chain for being carbon number between 2~4.
3. Fluorescent star-like polymer according to claim 1 or 2, it is characterised in that described Fluorescent star-like polymer is
Poly- ILE Methacrylamide, shown in chemical structural formula such as formula (V)：
Wherein, n=15.
4. the preparation method of Fluorescent star-like polymer described in claim 1, it is characterised in that monomeric compound 2 is in initiator
In the presence of compound 3, Fluorescent star-like polymer is obtained using ATRP living polymerizations；
Shown in the structure of monomeric compound 2 such as formula (II)：
Shown in the structure of compound 3 such as formula (III)：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number, k=1~4, m=1~3.
5. preparation method according to claim 4, it is characterised in that the consumption of the versus compound 3 of compound 2 is 10~
500 times, reacted under conditions of 60~70 DEG C.
6. preparation method according to claim 5, it is characterised in that compound 3 is dissolved in organic solvent so that chemical combination
The concentration of thing 3 adds after the CuBr with the amount of the grade material of compound 3 in the range of 2~5g/L, carries out 3~4 freezing-pump drainages
Circulation；Afterwards under nitrogen protection, the part PMDETA of the amount of 5~15 times of materials of compound 3 of addition and 10~500 times of changes
The monomeric compound 2 of the amount of the material of compound 3；Again after freezing pump drainage, stir 10~15 minutes, make CuBr and the complete network of part
Close, reacted 30~120 minutes at 60~70 DEG C；After reaction terminates, solvent is removed, is dialysed, freeze-drying.
7. preparation method according to claim 6, it is characterised in that the organic solvent is the mixing of butanone and methanol
The volume ratio of thing, butanone and methanol is 2:1.
8. the preparation method according to any one of claim 4~7, it is characterised in that the preparation side of the monomeric compound 2
Method is：Compound 1 is dissolved in sodium hydrate aqueous solution so that the pH value of solution is then added dropwise more than 10 at 0 DEG C
Methacrylic chloride, and supplement corresponding sodium hydroxide solution to maintain pH value between 9~11；After completion of dropping, in room temperature
1~3 hour of lower stirring, solution is acidified with watery hydrochloric acid afterwards so that pH value is down between 1~3；Extracted with ethyl acetate
Take, remove most of solvent with Rotary Evaporators, recrystallized by hot saturated solution method, obtain compound 2；
Shown in the structure of compound 1 such as formula (I)：
Wherein, R is hydrogen atom or the alkyl for being 1~9 containing carbon number.
9. Fluorescent star-like polymer described in any one of claims 1 to 3 is used as the application of protein molecular carrier.
10. application according to claim 9, it is characterised in that using Fluorescent star-like polymer as protein molecular carrier, take
Band external source toxalbumin molecule enters in intracellular or insect bodies.
11. application according to claim 10, it is characterised in that the Fluorescent star-like polymer is mixed with foreign protein molecule
Conjunction forms complex, is then incubated altogether with cell, the fluorescence star for making to carry foreign protein molecule by endocytosis
Polymer support enters intracellular, or the method by injecting or being fed with insect, makes the fluorescence star for carrying foreign protein molecule
Shape polymer support enters in insect bodies.
12. application according to claim 10, it is characterised in that the foreign protein molecule for positive charge histone or
The Bt toxalbumin of negative electrical charge.
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| CN108794710B (en) * | 2018-04-18 | 2019-10-18 | 中国农业大学 | A kind of star-type polymer and its preparation method and application |
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| CN1447843A (en) * | 2000-08-14 | 2003-10-08 | 马克思-普朗克科学促进协会公司 | Functionalized perylene tetracarboxylic acid diimides |
| CN103172776A (en) * | 2013-04-08 | 2013-06-26 | 中国农业大学 | Fluorescent star-like polymer as well as preparation method and application thereof |
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